柚皮苷通过调控TGF-β1/Smad2信号通路改善脓毒症急性肺损伤
Improvement of sepsis-related acute lung injury through Naringin by regulating TGF-β1/Smad2 signaling pathway
何萍萍 1邓宇 2王俞涵 1张臻 3王浩 3潘广涛4
作者信息
- 1. 湖北中医药大学黄家湖医院康复科,武汉 430065
- 2. 广州中医药大学附属广州市中西医结合医院中医科,广州 510800
- 3. 南京中医药大学附属盐城中医院呼吸科,盐城 224000
- 4. 南京中医药大学附属盐城中医院皮肤科,盐城 224000
- 折叠
摘要
目的 研究柚皮苷对脓毒症急性肺损伤模型大鼠的保护作用,并探讨其作用机制.方法 采用随机数字表法将30只雄性SD大鼠分为假手术组、模型组、柚皮苷低剂量组、柚皮苷中剂量组、柚皮苷高剂量组,每组6只.除假手术组外,其余各组均行盲肠结扎穿刺术建立脓毒症相关的急性肺损伤模型.造模后,柚皮苷低、中、高剂量组分别灌胃20、40、80 mg/kg柚皮苷,假手术组和模型组灌胃等体积蒸馏水,1次/d,给药2 d.采用HE染色观察肺组织病理学变化,ELISA检测肺泡灌洗液(BALF)中IL-1、IL-6、IL-18水平,免疫荧光法检测肺组织TNF-α表达,Western Blot检测肺组织TGF-β1、转化生长因子β受体Ⅰ(TGF-βR1)、Smad2蛋白表达.另设激动剂组、柚皮苷加激动剂组,每组6只,采用免疫组化染色检测各组肺组织TGF-β1、Smad2蛋白表达,验证柚皮苷对TGF-β1、Smad2蛋白表达的影响.结果 与模型组比较,柚皮苷各剂量组肺组织病理性损伤明显减轻,BALF中 IL-1β、IL-6、IL-18 水平降低(P＜0.01),肺组织 TNF-α 及 TGF-β1、TGF-βR1、Smad2 蛋白表达降低(P＜0.01或P＜0.05).进一步验证发现,激动剂组TGF-β1、Smad2免疫组化评分升高(P＜0.01),柚皮苷加激动剂组TGF-β1、Smad2蛋白表达降低(P＜0.01).结论 柚皮苷可减轻大鼠肺部炎症反应以保护脓毒症相关的急性肺损伤,其保护作用与抑制TGF-β1/Smad2信号通路有关.
Abstract
Objective To investigate the protective effect of naringenin on acute lung injury related with sepsis;To discuss its possible mechanism.Methods Totally 30 male SD rats were randomly divided into sham-operation group,model group,naringin low-,medium-and high-dosage groups,with 6 rats in each group.The sepsis-related acute lung injury model was established by cecal ligation and puncture in all groups except the sham-operation group.After modeling,naringin low-,medium-and high-dosage groups were given naringin 20 mg/kg,40 mg/kg and 80 mg/kg,respectively for gavage,while the sham-operation group and the model group were given the same volume of distilled water by gavage,once a day,for 2 days.Pathological changes in lung tissue were observed using HE staining.The levels of 1L-1,IL-6 and IL-18 in bronchoalveolar lavage fluid(BALF)were measured by ELISA;the expression of TNF-α in lung tissue was detected by immunofluorescence histopathology;the expressions of TGF-β1,TGF-βR1 and Smad2 were detected by Western Blot.An agonist group and a naringin plus agonist group were set up,with 6 mice in each group,and the expressions of TGF-β1 and Smad2 protein in the lung tissue of each group were detected by immunohistochemical staining to verify the effect of naringin on the expressions of TGF-β1 and Smad2 protein.Results Compared with the model group,the pathological injury of lung tissue in naringin groups were obviously alleviated,and the levels of IL-1β,IL-6 and IL-18 in BALF decreased(P＜0.01),the protein expressions of TNF-α,TGF-β1,TGF-βR1 and Smad2 in lung tissue decreased(P＜0.01 or P＜0.05).Further verification found that the expressions of TGF-β1 and Smad2 in the agonist group increased(P＜0.01),while the expressions of TGF-β1 and Smad2 in the naringin agonist group decreased(P＜0.01).Conclusion Naringin can reduce the inflammatory response in the lung of the rats to protect against sepsis-related acute lung injury,and its protective effect could be related to the inhibition of the TGF-β1/Smad2 signaling pathway.
关键词
柚皮苷/脓毒症/急性肺损伤/炎症/大鼠Key words
Naringin/Sepsis/Acute lung injury/Inflammation/Rats引用本文复制引用
基金项目
江苏省卫生健康委医学科研项目(Z2021068)
2021年盐城市医学科技发展计划重点项目(YK2021004)
2022年度江苏省科协青年科技人才托举工程(TJ-2022-097)
出版年
2024
NETL
NSTL
万方数据